Xylem sap residue in cut-open conduits can affect gas discharge in pneumatic experiments

Considerable attention has been paid to address methodological concerns related to measurements of embolism in conduits of angiosperm xylem. A fast, easy, and cheap method is based on gas extraction measurements from dehydrating samples to obtain pneumatic vulnerability curves (VCs). Here, we tested the assumption that cutting open conduits leads to gas-filled lumina when these are cut in air at fairly high water potentials, which is required to detect embolism in intact conduits. We performed VCs with the Pneumatron for 12 angiosperm species, and extracted sap from cut-open vessels in branches of nine species under early stages of branch dehydration. The optical method was applied to Citrus plants as an alternative reference method to estimate embolism resistance. We found an increase in gas discharge during early stages of dehydration, which affected the pneumatic VCs for most of the species studied. Xylem sap residue was not absorbed immediately by surrounding tissue in cut-open conduits in six of the nine species, but gradually disappeared over time during progressive dehydration. The amount of gas discharged increased until all residual sap was absorbed, and was not related to embolism. We conclude that residual xylem sap in cut-open conduits affects early stages of pneumatic VCs, and represents a novel artefact that can easily be corrected for. Yet, it remains unclear why exactly the air-water meniscus in cut-open conduits did not fully withdraw to the conduit end wall in most species. By analysing the slope of VCs over time, we could improve estimations of embolism resistance, as evidenced by a strong agreement between the pneumatic and the optical methods. Since residual sap in cut-open conduits of some species could slightly underestimate embolism resistance, we propose to apply a correction for this artefact based on the high time resolution measurements taken with a Pneumatron.